Determination of Hydrogen Peroxide by Fluorescein Chemiluminescence Catalyzed with Horseradish Peroxidase Encapsulated in Liposome

• KAMIDATE Tamio

  Division of Molecular Chemistry, Graduate School of Engineering, Hokkaido University

• ISHIDA Yoshiki

  Division of Molecular Chemistry, Graduate School of Engineering, Hokkaido University

• TANI Hirofumi

  Division of Molecular Chemistry, Graduate School of Engineering, Hokkaido University

• ISHIDA Akihiko

  Division of Molecular Chemistry, Graduate School of Engineering, Hokkaido University

Bibliographic Information

Other Title

• リポソームに内封したペルオキシダーゼを触媒に用いるフルオレセイン化学発光による過酸化水素の定量
• リポソーム ニ ナイ フウ シタ ペルオキシダーゼ オ ショクバイ ニ モチイル フルオレセイン カガク ハッコウ ニ ヨル カサンカスイソ ノ テイリョウ

Abstract

Fluorescein (FL) and H₂O₂ rapidly permeated into the inner phase of liposome which trapped horseradish peroxidase (HRP), to initiate HRP-catalyzed FL chemiluminescence (CL) with H₂O₂. The CL intensity was dependent on the concentration of H₂O₂. The optimum conditions of charge-type, composition and diameter of liposome in the assay of H₂O₂ were determined by measuring the CL intensity, to be maximal under optimum conditions. Anionic liposome containing phosphatidylcholine, dimyristoyl-glycero-phosphocholine and cholesterol (Chol) was effective for enhancing the CL intensity and stability of liposome. The CL intensity decreased with an increase in the content of Chol in liposome. The optimal content of Chol was thus determined to be 10 mol%. The effect of the liposome size on the CL intensity was examined by preparing liposomes with a different diameter. The CL intensity increased with an increase in the diameter of liposome. The optimal diameter of liposome was thus determined to be 1000 nm. The logarithmic calibration curve of H₂O₂ was linear over the range from the detection limit of 4.0 × 10⁻⁸ M up to 1.0 × 10⁻⁵ M. When HRP trapped in liposome was used as a catalyst, the CL intensity was greater than that observed by using HRP dissolved in the bulk solution in the range of 4.0 × 10⁻⁷ M up to 1.0 × 10⁻⁵ M of H₂O₂.<br>

Journal

• BUNSEKI KAGAKU

  BUNSEKI KAGAKU 54 (6), 569-572, 2005

  The Japan Society for Analytical Chemistry

Keywords

• chemiluminescence
• liposome
• peroxidase
• hydrogen peroxide
• fluoresein

Details

• CRID: 1390001204354912640
• NII Article ID: 110002910102
• NII Article ID: 120006531131
• NII Book ID: AN00222633
• DOI: 10.2116/bunsekikagaku.54.569
• COI: 1:CAS:528:DC%2BD2MXlsVSqsrg%3D
• HANDLE: 2115/71714
• NDL BIB ID: 7401000
• ISSN: 05251931
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I7401000
• Web Site: http://www.jstage.jst.go.jp/article/bunsekikagaku/54/6/54_6_569/_pdf
• Web Site: http://dl.ndl.go.jp/info:ndljp/pid/10896143
• Text Lang: ja
• Data Source:
  • JaLC
  • IRDB
  • NDL
  • Crossref
  • NDL-Digital
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed